OBJECTIVETo investigate the effect of midazolam on human ether-a-go-go (hERG) K+ channels exogenously expressed in human embryonic kidney cells (HEK-293) and the underlying molecular mechanisms.

METHODSWhole-cell patch clamp technique was used to record WT, Y652A and F656C hERG K+ current expressed in HEK-293 cells.

RESULTSMidazolam inhibited hERG K+ current in a concentration-dependent manner, the half-maximum block concentrations (IC50) values were (1.31 ± 0.32) µmol/L. The half-activation voltage (V1/2) were (2.32 ± 0.38) mV for the control and (-1.96 ± 0.83) mV for 1.0 µmol/L midazolam. The half-inactivation voltage (V1/2) was slightly shifted towards negative voltages from (-49.25 ± 0.69) mV in control to (-57.53 ± 0.53) mV after 1.0 µmol/L midazolam (P < 0.05). Mutations in drug-binding sites (Y652A or F656C) of the hERG channel significantly attenuated the hERG current blockade by midazolam.

CONCLUSIONMidazolam can block hERG K+ channel and cause the speed of inactivation faster. Mutations in the drug-binding sites (Y652 or F656) of the hERG channel were found to attenuate hERG current blockage by midazolam.

Dose-Response Relationship, Drug ; Ether-A-Go-Go Potassium Channels ; drug effects ; HEK293 Cells ; Humans ; Midazolam ; pharmacology ; Mutation ; Patch-Clamp Techniques ; Potassium Channel Blockers ; pharmacology

BACKGROUND:The adhesion between muscle tendon and surrounding tissue after tendon restoration is one hot topic in clinic.
OBJECTIVE:To construct a deep flexor tendon transection model in the third toe of female Leghorn chicken and to explore the effect of peritenon transplantation on the prevention of tendon adhesions.
METHODS:After the transection models were successful y established, the third toe of left claw was taken as experimental group A. The transected tendon was sutured, the tendon anastomosis end was wrapped with the peritenon of the deep flexor tendon from the ipsilateral fourth toe (experimental group B). The third toe of right claw served as control group A, the transfected tendon was sutured and restored with the peritenon. The fourth toe of right claw was taken as control group B. Gross observation and histological observation of the tendon were performed.
RESULTS AND CONCLUSION:At 28 days postoperatively, gross observation and histological observation of Leghorn chicken were performed and compared using the Kruskal-Wal isH and Nemenyi test, respectively. The results showed that, the therapeutic effect was better in experimental group A than in control group A (P<0.05), but slightly poorer compared with control group B and experimental group B (P<0.05). The postoperative effect was better in experimental group B than in control group A (P<0.05) and showed no significant difference between experimental group B and control group B (P>0.05). The flexor function was evaluated and compared with the least significant difference t-test. The results showed that the postoperative effect was better in experimental group A than in control group A (P<0.05), but slightly poorer compared with control group B and experimental group B (P<0.05). The postoperative effect was better in experimental group B than in control group A (P<0.05) and showed no significant difference compared with control group B (P>0.05). Peritenon transplantation can effectively prevent tendon adhesions and has little impact on normal tendon sliding.

BACKGROUND:Tendon adhesions after tendon repair have been one of the problems difficult to solve in clinical practice. OBJECTIVE:To introduce the current situation and progress of the prevention of postoperative tendon adhesions. METHODS: A computer-based search of CNKI, Wanfang, PubMed databases for articles related to prevention of tendon adhesions published from January 1971 to May 2014 using the keywords of “tendon injury; tendon adhesions; progress” in Chinese and English, respectively. Repetitive content and irrelevant articles were excluded, and finaly 40 articles were included in result analysis. RESULTS AND CONCLUSION: In this paper, we analyze the characteristics of tendons and causes of tendon adhesions, summarize the methods for prevention of tendon adhesions and the therapeutic effects, and thereby analyze the pros and cons of various methods and materials for prevention of tendon adhesions. Prevention of tendon adhesions after tendon repair involves various aspects: thorough debridement, better surgical approach, microsurgical repair of tendon tissues, applications of drugs and barrier materials, physical therapy as wel as appropriate functional exercise after tendon repair. The integrated use of these means can be done to prevent the occurrence of postoperative tendon adhesions.

BACKGROUND: According to present theories and our clinical experience, immune ablative and tolerance inducing theory is proposed. Immune ablative means to clear out mutate cell clones and without transfusion of hemopoietic stem cells afterwards; intolerance inducing means to induce animal models not to react to mutate somatic cells, which avoids relapse or new occurrence of autoimmune disease. OBJECTIVE: To explore the effects of immune-ablative and tolerance inducing therapy in treating animal model of immune polymyositis (PM). DESIGN, TIME AND SETTING: Randomized, controlled animal experiment was performed at the Animal Experimental Center of Nanfang Hospital from December 2008 to April 2009. MATERIALS: One New Zealand rabbit, female, weighing 4.1 kg and 36 England guinea pigs, female, weighing 400-500 g, were used. METHODS: New Zealand rabbit's muscle tissue homogenate and complete Freund's adjuvant (CFA) were injected into guinea pigs to make PM animal models. The 28 animal models were randomly divided into intense immune-ablative and tolerance inducing group (Busulfan 1 mg/kg, every 12 hours, totally 8 doses; followed by CTX 40 mg/kg per day for 4 days; then cyclosporine A (CsA) 3 mg/kg per day was given till animals were dead); cyclophosphamide (CTX) group: CTX was given, 10 mg/kg per day for 3days; immune-ablative and tolerance inducing group: Busulfan 0.8 mg/kg, CTX 30 mg/kg, CsA 3 mg/kg; the administration time and dose were the same as group 1. Control group was not treated.MAIN OUTCOME MEASURES: Full blood count (FBC) and biochemical index were tested before and after treatment, and surviving time was recorded. In addition, muscle pathological changes were observed.RESULTS: Compared with control group, number of white cells was significantly decreased in the other groups, and hematopoiesis function gradually restored after administration. The number of white cells in the immune-ablative and tolerance inducing group was the most, and striated muscle pathology showed PM. Following administration, the glutamic oxaloacetic transaminase and creatine kinase of intense immune-ablative and tolerance inducing and immune-ablative and tolerance inducing groups were significantly reduced (P < 0.05, P < 0.01), but no obvious striated muscle pathological changes were found. The glutamic oxaloacetic transaminase, lactic dehydrogenase and creatine kinase in the CTX and control groups remained unchanged. Survival time of intense immune-ablative and tolerance inducing group was the shortest among all groups, and there was no significant difference between CTX and control groups. The animals in immune-ablative and tolerance inducing group survived for the longest time. CONCLUSION: Immune-ablative and tolerance inducing therapy has preferable effect on treating animal models of PM, and its prognosis is better than intense immune-ablative and tolerance inducing therapy and regular CTX therapy.

Objective To use CT and 3D reconstruction technique to locating mark of sustentaculum tali ,and measure the an‐gle & length of the specified point of the lateral wall of the calcaneus to sustentaculum tali for providing .To basis for the operation of calcaneus .Methods Thirty adult ankle wet specimens were chosen and scanned with CT ,and the dicom data were converted into the software to 3D‐reconstruction ,the observation and measurement to find new methods of locating mark of sustentaculum tali ;Forty adult calcaneus dried specimens were chosen and scanned with CT .The dicom data were converted into the software to 3D‐re‐construction ,using a workstation software measure the angle and length of the line which from the center of the sustentaculum tail to each point on the calcaneal lateral wall .Results First ,in the function of ankle joint position ,with the foot medial surface as “sag‐ittal reference plane” measurement ,the vertical line passing through the leading edge of medial malleols and the coronal line passing through the leading edge of sustentaculum tali intersect at a point .The vertical distance of the point to leading edge of medial malle‐ols was (34 .95 ± 2 .60) mm ,the length of sustentaculum tali was (21 .44 ± 1 .89) mm ,the height of sustentaculum tali was (10 .17 ± 1 .16) mm ,the anteversion of sustentaculum tali was (35 .31 ± 3 .73)° .Second ,in front of calcaneus ,upward angel of points G ,E were (28 .78 ± 3 .90 )° ,(29 .47 ± 2 .96 )° respectively .Frontward angle of points G ,E were ( - 19 .83 ± 4 .60 )° , ( - 11 .02 ± 4 .74)° respectively .Under the posterior articular surface ,upward angel of points C ,K ,I ,J were(2 .52 ± 2 .46)° ,(2 .92 ± 2 .28)° ,(14 .98 ± 2 .49)° ,(14 .38 ± 1 .90)° respectively .Frontward angle of points C ,K ,I ,J were(7 .32 ± 1 .66)° ,(19 .25 ± 1 .98)° , (10 .30 ± 2 .63)° ,(19 .33 ± 1 .97)° respectively .The length of screw was about 40 - 44 mm .The length of C point to the sustentacu‐lum tail was minimum ,and the length of G was maximum .Conclusion In the function of ankle joint position ,the sustentaculum tail can be located with the leading edge of medial malleols as a surface landmarks .The measurement of the angle and length of which from each point on the calcaneal lateral wall to sustentaculum tail by using CT .

Objective To analyze the clinical characteristics,treatment and prognosis of primary lymphoma of bone (PLB).Methods Retrospective analysis of 9 cases with newly diagnosed PLB was performed.The primary lesion was located in bone and no evidence of lesion was found in other tissues.All the lesions were confirmed by histological examination and immunohistochemical staining.The clinical manifestation and radiological appearance of all cases were non-specific.Results 9 patients were enrolled in the current study including 6 males and 3 females with average age of 34 years old (15-71 years old).All cases had suffered local pain or edema as initial symptom.Single bone infiltration was detected in 7 patients,while multiple bones infiltration was noticed in 2 patients.The lesion distribution was as follows:4 lesions were situated in pelvis (3 in ilium and 1 in sacral),1 in femur,1 in humerus,1 in scapula,and 2 in spine.2 cases were treated with radiotherapy and chemotherapy,3 cases were treated with chemotherapy and hematopoietic stem cell transplantation,1 case was treated with chemotherapy and surgery,and 3 cases received chemotherapy alone.With in a median follow-up of 8 months,6 patients showed overall response (including 5 cases of complete response and 1 case of partial response).Conclusions PLB is a rare condition,and the diagnostic criteria and treatment are still not well defined.Early diagnosis and prompt treatment usually achieves satisfactory prognosis.The possibility of PLB must be concerned for patients with bone pain,soft tissue swelling or unexplained fractures,and pathological examination should be carried on as soon as possible.

Serum cortisol and plasma ACTH were determined in 10 patients with severe sepsis and 12 with septic shock on day 1,3,5 after diagnosis were made,and the data were compared with 12 control patients. To evaluate the hypothalamic-pituitary-adrenal(HPA)axis function in patients with severe sepsis and septic shock,1?g ACTH stimulation test was applied after hormone concentrations were obtained on day 1.Compared with the control patients,ACTH level was significantly higher in patients with severe sepsis and lower in septic shock(P

Pyrimidine derivatives have been the subject of much attention in pesticide and medicine fields owing to their unique biological properties. Particularly, a large number of these compounds have recently been reported to show substantial antitumor activities, and some of them have been investigated in clinical trials. Although these structurally novel compounds have a common chemical moiety of a pyrimidine ring, there are a variety of mechanisms of their antitumor action, such as, inhibition of cyclin-dependent-kinases, inhibition of protein tyrosine kinase, inhibition of carbonic anhydrases, inhibition of dihydrofolate reductase and disruption of microtubule assembly. In this paper, we described the latest advances in the research of such pyrimidine derivatives as antitumor drug according to their action on targets.
Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; therapeutic use ; Carbonic Anhydrase Inhibitors ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinases ; antagonists & inhibitors ; Folic Acid Antagonists ; pharmacology ; Humans ; Neoplasms ; drug therapy ; pathology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; chemistry ; pharmacology ; therapeutic use ; Tetrahydrofolate Dehydrogenase ; pharmacology ; Tubulin Modulators ; pharmacology ; therapeutic use

BACKGROUNDThere are few researches for the healing of metaphyseal fractures; moreover, the animal models to study the metaphyseal fractures are usually made by the oscillating saw osteotomy without reliable fixation, which is not in accordance with our current clinical practice. In this study, we established a new model to observe the healing process of metaphyseal fractures.

METHODSEighteen New Zealand rabbits were used in the study. The fracture model was created by splitting the medial tibial plateau in rabbits, then reset, and fixed with compression screws. At 1, 2, 3, 4, 6, and 8 weeks postoperatively, the tibial specimens were collected; firstly, a general observation and an X-ray examination of the specimens was done, and then they were embedded in methylmethacrylate and cut into sections with hard tissue slicer. The sections were stained with Giemsa reagent and examined under light microscopy.

RESULTSThere was no fracture displacement in the tibial specimens of all time points, except for one showing a collapse. No external callus formation could be observed by X-ray and general examination. After 1 week of the operation, the fracture gap was filled by mesenchymal tissue; 2 weeks postoperatively, a large number of woven bones were formed; from the third week onwards, the woven bone began to turn into lamellar bone, and new trabecular structure began to form. In all of the slices, no obvious chondrocytes formed in fracture areas; thus, there was no endochondral ossification.

CONCLUSIONSThis model was an ideal fracture animal model and suitable for the study of metaphyseal fracture healing. The X-ray and histological images demonstrated that metaphyseal fracture healing was a process of direct bone healing through intramembranous bone formation under the conditions of minor trauma, good reduction, and firm fixation.

Animals ; Fracture Healing ; physiology ; Fractures, Bone ; diagnostic imaging ; pathology ; Rabbits ; Radiography

OBJECTIVETo investigate the role and significance of Wnt/beta-catenin signaling pathway regulating GSK-3beta, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC).

METHODSThe HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against beta-catenin. Proteins were extracted and the expressions of beta-catenin, GSK-3beta, p-GSK-3beta, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection.

RESULTSbeta-catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P < 0.05). Interestingly, GSK-3beta and p-GSK-3beta expressions increased gradually at 72 and 96 h (tGSK-3beta= 4.98, tp-GSK-3beta= 29.83, P < 0.05) respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P > 0.05). Smad3 expression was increased at 72 h (t = 10.67, P < 0.05) and decreased to normal at 96 h (t = 1.26, P < 0.05), while TERT expression decreased at 72 h (t = 4.18, P is less than 0.05) and increased to normal at 96 h (t = 1.26, P > 0.05).

CONCLUSIONSWnt/beta-catenin signaling pathway is related to the expressions of GSK-3beta, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/beta-catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Small Interfering ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism